Ellis J A, Fish W R, Sileghem M, McOdimba F
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
Vet Parasitol. 1993 Oct;50(1-2):143-9. doi: 10.1016/0304-4017(93)90015-f.
We have adapted a tetrazolium salt (MTT) colorimetric cytotoxicity assay to the assessment of viability and metabolic function in cultured African trypanosomes. Trypomastigotes of Trypanosoma congolense and T. brucei rhodesianse were harvested from the blood of parasitemic rats and cultured under axenic conditions that support trypanosome viability and growth. Analysis of serial dilutions of these bloodstream forms indicated that the assay could detect 10(4) parasites. To assess the effect of lymphoid cytokines on trypomastigote viability, 10(5) freshly harvested parasites were cultured with a wide range of dilutions of human recombinant IL-1, IL-3, IL-6, interferon-gamma (IFN gamma) or tumor necrosis factor-alpha (TNF alpha), or bovine recombinant IFN gamma or TNF alpha for 24, 48 or 96 h. These cytokines had no apparent growth enhancing or inhibitory effect on the trypomastigotes compared with growth in supplemented medium alone. This assay has several advantages over traditional counting methods, including increased sensitivity and rapid, repeatable quantitation. This adaptation of the MTT colorimetric assay should be useful in screening drugs and host-derived factors for growth-modulating effects on trypanosomes and other extracellular protozoan parasites.
我们已将一种四唑盐（MTT）比色细胞毒性测定法应用于培养的非洲锥虫的活力和代谢功能评估。从患寄生虫血症大鼠的血液中采集刚果锥虫和布氏罗得西亚锥虫的锥鞭毛体，并在支持锥虫活力和生长的无菌条件下进行培养。对这些血液形式的系列稀释液进行分析表明，该测定法可检测到10⁴个寄生虫。为了评估淋巴因子对锥鞭毛体活力的影响，将10⁵个 freshly harvested parasites与一系列稀释度的人重组白细胞介素-1（IL-1）、IL-3、IL-6、干扰素-γ（IFNγ）或肿瘤坏死因子-α（TNFα），或牛重组IFNγ或TNFα一起培养24、48或96小时。与仅在补充培养基中生长相比，这些细胞因子对锥鞭毛体没有明显的生长促进或抑制作用。该测定法与传统计数方法相比具有几个优点，包括更高的灵敏度以及快速、可重复的定量。MTT比色测定法的这种应用在筛选对锥虫和其他细胞外原生动物寄生虫具有生长调节作用的药物和宿主衍生因子方面应该是有用的。 （注：“freshly harvested parasites”这里可能是“刚收获的寄生虫”，但“freshly”在句中稍显突兀，可能原文表述有误，你可再确认下。）
